Related Experiment Video
Updated: May 13, 2026
Production and Purification of Non Replicative Canine Adenovirus Type 2 Derived Vectors
Published on: December 3, 2013
Transducing properties of a pre-structured α-helical DPT-peptide containing a short canine adenovirus type 2 E4orf4
A Galioot1, A N Godet, V Maire
1Unité Signalisation Moléculaire et Activation Cellulaire, Institut Pasteur, Paris, France.
Background:
Induction of the death pathway resulting from the specific interaction of the PP2A1 phosphatase with adenoviral E4orf4 protein is a promising approach for cancer therapy. With the aim of deregulating tumor pathways, and mimicking E4orf4 anti-cancer signal, we have previously proposed the DPT technology concept, based on design of specific PP1/PP2A interacting penetrating peptides.
Methods:
Using biochemical, structural and cell survival experiments, we have characterized new DPT-peptides containing short PP2A binding sequences.
Results:
We identified overlapping sequences, located within the N-terminal domain E4orf423-46 of canine adenoviral E4orf4 protein, that interact with the PP2A-Bα subunit of PP2A1 holoenzyme. We characterized DPT-E4orf44 and TAT-E4orf44, two bi-partite cell penetrating peptides containing the 12 PP2A1 binding residues of the canine type 2 E4orf427-38 sequence, respectively fused to the DPT-sh1 and TAT shuttle sequences. Surprisingly DPT-E4orf44, in contrast to inactive TAT-E4orf44, adopted a well defined α-helical structure and co-precipitated PP2A1 from HeLa cell extracts. DPT-E4orf44 also internalized streptavidin-HRP and inhibited survival of HeLa cells more efficiently than TAT, TAT-E4orf44 or the previously published anti-tumor TAT-derived peptide shepherdin. DPT-E4orf44 also efficiently inhibited the survival of human adherent transformed cells, including wild type and p53 mutated colonic HCT116 cells, without affecting survival of human non-transformed fibroblasts.
Conclusions:
We characterized the transducing properties of a new α-helical DPT-E4orf44 peptide containing a short PP2A-interacting sequence from canine Adenoviral E4orf4 protein.
General Significance:
Our results suggest that α-helical structured DPT peptides specifically interacting with PP2A could be a valuable anti-cancer drug design scaffold.
Insights
New DPT-E4orf44 peptides mimic adenoviral E4orf4 protein's anti-cancer signals by interacting with PP2A1 phosphatase. These peptides show potential as a scaffold for novel anti-cancer drug design.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- Adenoviral E4orf4 protein interaction with PP2A1 phosphatase can induce cancer cell death pathways.
- Developing therapies that mimic E4orf4's anti-cancer signaling is a promising strategy.
- The DPT (Drug Penetrating Technology) concept utilizes peptides designed to interact with PP1/PP2A phosphatases.
Purpose of the Study:
- To characterize new DPT-peptides designed to mimic the anti-cancer effects of adenoviral E4orf4.
- To investigate the interaction of these peptides with PP2A1 phosphatase and their anti-cancer activity.
Main Methods:
- Biochemical assays to study protein interactions.
- Structural analysis to determine peptide conformation.
- Cell survival experiments to assess anti-cancer efficacy.
- Characterization of cell-penetrating peptides (DPT-E4orf44 and TAT-E4orf44).
Main Results:
- Identified specific PP2A-binding sequences within the adenoviral E4orf4 N-terminal domain.
- DPT-E4orf44, an α-helical peptide, effectively co-precipitated PP2A1 and inhibited cancer cell survival.
- DPT-E4orf44 demonstrated superior internalization and anti-cancer activity compared to control peptides in various cancer cell lines, including HCT116.
- Non-transformed fibroblasts remained unaffected by DPT-E4orf44 treatment.
Conclusions:
- Characterized the transducing properties and PP2A-interacting capabilities of the novel DPT-E4orf44 peptide.
- The α-helical structure and specific PP2A interaction of DPT peptides make them a promising scaffold for anti-cancer drug development.
Related Concept Videos
02:28Leaky Scanning
01:26Insertion of Single-pass Transmembrane Proteins in the RER
Integral transmembrane proteins possess transmembrane and extra membrane domains. The transmembrane domains are primarily made of 20-25 hydrophobic amino acids arranged in a helical secondary confirmation. These...